2023 Fiscal Year Final Research Report
Molecular mechanisms of onset and exacerbation of non-obstructive hydrocephalus caused by abnormal ventricular ependymal ciliary dynein
| Project/Area Number |
20K17959
|
|---|
| Research Category |
Grant-in-Aid for Early-Career Scientists
|
| Allocation Type | Multi-year Fund |
|---|
| Review Section |
Basic Section 56010:Neurosurgery-related
|
|---|
| Research Institution | Nagoya University |
|---|
Principal Investigator |
|
|---|
| Project Period (FY) |
2020-04-01 – 2023-03-31
|
| Keywords | 脳室繊毛 / ダイニン / クライオ電子顕微鏡 |
|---|
| Outline of Final Research Achievements |
Dpcd KO mice were used to analyze abnormal ciliary movements. We found that the amplitude of cilia movement was abnormal, which is observed in abnormal inner arm dynein. Next, immunostaining and RNA expression analysis of the inner arm dynein subunits revealed that the expression of Dnah6 was mainly downregulated. Furthermore, electron microscopic analysis of the internal structure showed that a portion of the inner arm dynein was present, confirming that it was not completely absent. Next, we attempted to isolate ventricular cilia. We succeeded in purifying an aqueous solution of cilia by using a concentration gradient, because shear stress is the most efficient way to recover cilia. The concentration of this aqueous cilia solution enabled 3D analysis by cryo-EM.
|
| Free Research Field |
脳神経外科
|
| Academic Significance and Societal Importance of the Research Achievements |
これまでDpcdKOによる繊毛運動異常は内腕ダイニンの欠損によるものと報告されていた。我々の研究の結果、内腕ダイニンは完全欠損しておらず一部の発現が低下していることが示唆された。さらに内部構造の把握を進めるため、脳室繊毛の単離に取り組んだ。これまでに脳室繊毛の単離に成功した報告はなく極めて新規性が高いものである。我々は内部構造把握にはクライオ電子顕微鏡による３D画像の取得が重要と考え、これに耐える濃度の繊毛水溶液作成に取り組んだ。その結果、十分な濃度の繊毛水溶液を作成し、クライオ電子顕微鏡観察に成功している。今後繊毛運動に由来する脳疾患の研究に役立つものと期待される。
|
